EP1940874B1 - Der botulinus neurotoxin a proteinrezeptor und seine anwendungen - Google Patents

Der botulinus neurotoxin a proteinrezeptor und seine anwendungen Download PDF

Info

Publication number: EP1940874B1
Authority: EP; European Patent Office
Prior art keywords: bont; polypeptide; homo sapiens; sv2c; antibody
Prior art date: 2005-10-28
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.): Not-in-force

Application number

EP06806613A

Other languages

German (de)

English (en)

French (fr)

Other versions

EP1940874A2 (de

Inventor

Andreas Rummel

Thomas Binz

Stefan Dr. MAHRHOLD

Johannes Wilhelm Prof. Dr. BIGALKE

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Toxogen GmbH

Original Assignee

Toxogen GmbH

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

2005-10-28

Filing date

2006-10-30

Publication date

2011-12-28

2006-10-30 Application filed by Toxogen GmbH filed Critical Toxogen GmbH

2008-07-09 Publication of EP1940874A2 publication Critical patent/EP1940874A2/de

2011-12-28 Application granted granted Critical

2011-12-28 Publication of EP1940874B1 publication Critical patent/EP1940874B1/de

Status Not-in-force legal-status Critical Current

2026-10-30 Anticipated expiration legal-status Critical

Links

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- C07K14/70571—Receptors; Cell surface antigens; Cell surface determinants for neuromediators, e.g. serotonin receptor, dopamine receptor
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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Definitions

BoNT / A to BoNT / G The anaerobic, gram-positive bacterium C. botulinum produces seven different serotypes of clostridial neurotoxins. These are referred to as the botulinum neurotoxins (BoNT / A to BoNT / G). Of these, BoNT / A and BoNT / B in particular cause a neuroparalytic disorder in humans and animals called botulism.
the spores of C. Botulinum are found in the soil, but may develop into improperly sterilized and sealed home-made canned foods, which are the cause of many botulism cases.
the present invention now introduces synaptic vesicle protein 2C (SV2C) as a receptor for BoNT / A.
SV2C synaptic vesicle protein 2C
the luminal domains of the proteins synaptophysin, synaptoporin, synaptogyrin I & III, SV2A, SV2B and SV2C were subcloned and recombinantly expressed and isolated in E. coli as the glutathione-S-transferase (GST) fusion protein.
GST glutathione-S-transferase
the H C fragments of the seven BoNTs and TeNT were both recombinantly expressed in E. coli expressed as well as translated with 35 S-methionine in vitro .
the affinity of the H C fragments to the luminal domains of the GST fusion proteins listed above was determined in glutathione-S-transferase (GST) pull-down experiments.
GST-SV2C derivatives the inhibition of neurotoxicity of BoNT / A and BoNT / B was analyzed on the isolated mouse nerve-muscle preparation (HDI), which is the physiological target of clostridial neurotoxins.
the preferential blocking of cholinergic transmission can be explained by the HC entering the neuron at the periphery. Central synapses are protected by the blood-brain barrier, which can not be overcome by proteins.
GST fusion proteins (0.15 nmol each) immobilized on 10 ⁇ l of GT-Sepharose beads were incubated with H c fragments (0.1 nmol) in the absence or presence of a bovine brain ganglioside mixture (18% GM1 , 55% GD1a, 10% GT1b, 2% other ganglosides, Calbiochem, 20 ⁇ g each) in a total volume of 100 ⁇ l Tris-NaCl-Triton buffer for 3 h at 4 ° C. The beads were collected by centrifugation, the supernatant removed and the separated beads washed three times each with 160 ⁇ l of the same buffer. The washed pellet fractions were boiled in SDS sample buffer and assayed with the supernatant fractions by SDS-PAGE and Coomassie blue staining, autoradiography or immunoblotting.
the luminal domains and the carboxyl-terminal transmembrane domains of the proteins synaptophysin, synaptoporin, synaptogyrin 1 & III, synaptotagmin II, SV2A, SV2B and SV2C were subcloned and expressed recombinantly in E. coli as glutathione-S-transferase (GST) fusion protein and isolated.
GST glutathione-S-transferase
the H c fragments of the seven BoNTs and tetanus neurotoxin (TeNT) were both recombinantly expressed in E. coli and translated in vitro with 35 S-methionine.

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EP06806613A 2005-10-28 2006-10-30 Der botulinus neurotoxin a proteinrezeptor und seine anwendungen Not-in-force EP1940874B1 (de)

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DE102005051789.7A DE102005051789B4 (de)	2005-10-28	2005-10-28	Der Botulinus Neurotoxin A Proteinrezeptor und seine Anwendungen
PCT/EP2006/010420 WO2007048638A2 (de)	2005-10-28	2006-10-30	Der botulinus neurotoxin a proteinrezeptor und seine anwendungen

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EP1940874A2 EP1940874A2 (de)	2008-07-09
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US (1)	US8476024B2 (ja)
EP (1)	EP1940874B1 (ja)
JP (1)	JP2009513118A (ja)
KR (1)	KR20080068069A (ja)
AT (1)	ATE539088T1 (ja)
AU (1)	AU2006308122B2 (ja)
CA (1)	CA2627457C (ja)
DE (1)	DE102005051789B4 (ja)
ES (1)	ES2379513T3 (ja)
NZ (1)	NZ567312A (ja)
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DE102004043009A1 (de)	2004-09-06	2006-03-23	Toxogen Gmbh	Transportprotein zum Einbringen chemischer Verbindungen in Nervenzellen
DE102005019302A1 (de)	2005-04-26	2006-11-16	Toxogen Gmbh	Carrier zum Targeting von Nervenzellen
US7985554B2 (en)	2005-10-14	2011-07-26	Wisconsin Alumni Research Foundation	Botulinum neurotoxin A receptor and the use thereof
EP2134749B1 (en)	2007-03-22	2013-11-06	The Regents of the University of California	Therapeutic monoclonal antibodies that neutralize botulinum neurotoxins
WO2010014854A2 (en)	2008-07-31	2010-02-04	The Regents Of The University Of California	Antibodies that neutralize botulinum neurotoxins
US8771707B2 (en) *	2008-09-30	2014-07-08	Wisconsin Alumni Research Foundation	Botulinum neurotoxin E receptors and uses thereof
WO2012047427A2 (en)	2010-08-31	2012-04-12	The Regents Of The University Of California	Antibodies for botulinum neurotoxins
WO2013180799A1 (en)	2012-05-30	2013-12-05	President And Fellows Of Harvard College	Engineered botulinum neurotoxin
BR112015003591B1 (pt) *	2012-11-21	2022-02-01	Ipsen Bioinnovation Limited	Usos de lys-c e métodos para fabricação de um polipeptídeo proteoliticamente processado
EP3274364B1 (en)	2015-03-26	2021-08-04	President and Fellows of Harvard College	Engineered botulinum neurotoxin
KR101670188B1 (ko)	2015-11-09	2016-10-27	충북대학교 산학협력단	보툴리눔 신경독소 ｅ형에 특이적인 폴리펩타이드 및 이의 용도
US11117935B2 (en)	2016-08-24	2021-09-14	President And Fellows Of Harvard College	Engineered botulinum neurotoxin
WO2018053004A2 (en)	2016-09-13	2018-03-22	Allergan, Inc.	Non-protein clostridial toxin compositions
KR101958312B1 (ko) *	2017-09-06	2019-07-02	국방과학연구소	보툴리눔 a형 독소에 대한 항체
IL272002A (en) *	2020-01-13	2021-07-29	The Israel Institute Of Biological Res Iibr	Methods for identifying active compounds against Clostridium neurotoxin
AU2023259903A1 (en) *	2022-04-25	2024-11-21	Jiksak Bioengineering, Inc.	Targeting agent

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US20070135514A1 (en) *	2002-12-03	2007-06-14	Berkley Lynch	Methods for the identification of agents for the treatment of seizures, neurological diseases, endocrinopathies and hormonal diseases
US7985554B2 (en) *	2005-10-14	2011-07-26	Wisconsin Alumni Research Foundation	Botulinum neurotoxin A receptor and the use thereof

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- 2005-10-28 DE DE102005051789.7A patent/DE102005051789B4/de not_active Expired - Fee Related
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AU2006308122A1 (en)	2007-05-03
JP2009513118A (ja)	2009-04-02
KR20080068069A (ko)	2008-07-22
CA2627457A1 (en)	2007-05-03
CA2627457C (en)	2014-05-13
US20090252722A1 (en)	2009-10-08
WO2007048638A3 (de)	2007-07-26
US8476024B2 (en)	2013-07-02
ZA200803403B (en)	2009-02-25
AU2006308122B2 (en)	2011-06-30
ATE539088T1 (de)	2012-01-15
ES2379513T3 (es)	2012-04-26
EP1940874A2 (de)	2008-07-09
NZ567312A (en)	2011-09-30
WO2007048638A2 (de)	2007-05-03
DE102005051789A1 (de)	2007-05-10
DE102005051789B4 (de)	2014-08-07

Publication	Publication Date	Title
EP1940874B1 (de)	2011-12-28	Der botulinus neurotoxin a proteinrezeptor und seine anwendungen
EP3181578B1 (de)	2019-01-30	Carrier zum targeting von nervenzellen
DE60032367T3 (de)	2011-03-10	Aktivierbare rekombinante neurotoxine
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Rummel et al.	2009	Botulinum neurotoxins C, E and F bind gangliosides via a conserved binding site prior to stimulation‐dependent uptake with botulinum neurotoxin F utilising the three isoforms of SV2 as second receptor
EP1786832B1 (de)	2012-07-18	Transportprotein zum einbringen chemischer verbindungen in nervenzellen
DE69233157T2 (de)	2004-05-19	Rekombinanter thrombinrezeptor und verwandte arzneimittel
DE60030429T2 (de)	2007-10-31	Neuronale, die exozytose hemmende peptide und diese enthaltende kosmetische und pharmazeutische zusammensetzungen
Leveque et al.	1994	Purification of the N-type calcium channel associated with syntaxin and synaptotagmin. A complex implicated in synaptic vesicle exocytosis.
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DE69434067T2 (de)	2006-03-02	Menschliche neuronale nikotin-acetylcholin-rezeptor-verbindungen und methoden ihres einsatzes
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DE69427740T2 (de)	2002-05-23	Inhibitoren einer Protease, die den löslichen p55-TNF-Rezeptor vom zellgebundenen TNF-Rezeptor abspaltet
Wellhöner	1992	Tetanus and botulinum neurotoxins
DE60123635T2 (de)	2007-08-16	Methode und zusammensetzung zur förderung von osteogenese
DE69434704T2 (de)	2006-10-19	Menschliches erythropoietin-rezeptor-fragment und antikörper dafür
Poulain et al.	1989	Multiple domains of botulinum neurotoxin contribute to its inhibition of transmitter release in Aplysia neurons
Khvotchev et al.	2000	α‐latrotoxin triggers transmitter release via direct insertion into the presynaptic plasma membrane
Apland et al.	2003	Inhibition of neurotransmitter release by peptides that mimic the N-terminal domain of SNAP-25
US6753315B2 (en)	2004-06-22	α-bungarotoxin molecules and uses thereof
EP1319022A2 (de)	2003-06-18	Breit einsetzbares verfahren zur identifizierung von modulatoren von g-protein gekoppelten rezeptoren
Beecher et al.	1997	Botulinum neurotoxin type A: Limited proteolysis by endoproteinase Glu-C and α-chymotrypsin enhanced following reduction; Identification of the cleaved sites and fragments
DE69932351T2 (de)	2007-07-05	DNS-kodierende Fusionsproteine, die durch Kupfer(II)-Ionen spezifisch spaltbar sind
AT412514B (de)	2005-03-25	Verfahren zum screenen von npt2b-modulierenden mitteln
DE19808258A1 (de)	1998-09-03	Kopfaktivator-bindendes Protein oder Polypeptid und Verfahren zur Suche nach funktionalen Agonisten oder Antagonisten unter Verwendung von kopfaktivatorbindendem Protein oder Polypeptid

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